Chemometric Estimation of the Contributions of Metals and Volatile Compounds to the Sensory Properties of Some Natural Grape Wines

Journal of Analytical Chemistry - Based on the concentrations of metals and volatile compounds in red and white wine samples and using linear and general linear models, we performed a comparative...     

Volatile compounds and odor preferences of ground beef added with garlic and red wine,and irradiated with charcoal pack

Irradiation is the most efficient non-thermal technology for improving hygienic quality and extending the shelf-life of food products. One of the adverse effects of food irradiation, however, is off-flavor production, which significantly affects the sensory preferences for certain foods. In this study, garlic (5%, w/w) and red wine (1:1, w/w) were added to ground beef to increase the radiation sensitivity of pathogens and improve meat odor/flavor. Samples were irradiated at 0 or 5 kGy in the presence of charcoal pack. SPME-GC–MS analysis was performed to measure the changes in the volatile compounds and sensory characteristics of the samples. The amount of total volatile compounds produced from ground beef was greater when the sample was irradiated. When garlic and red wine were added to the ground beef, the amount of volatile compounds significantly increased, and the amount of volatile compounds increased even further after irradiation. However, when the samples were irradiated with charcoal pack, the amount of volatile compounds decreased significantly. Sensory evaluation indicated that charcoal pack significantly increased the odor preferences for both irradiated and non-irradiated ground beef added with garlic. These results indicated that addition of charcoal pack to ground beef could reduce off-odor problems induced by irradiation, and this effect was consistent even when certain additives such as garlic and red wine were added.     

Stable isotope dilution analysis of wine fermentation products by HS-SPME-GC-MS

The aim of this study was to quantify, in a single analysis, 31 volatile fermentation-derived products that contribute to the aroma of red and white wine. We developed a multi-component method based on headspace solid-phase microextraction coupled with gas chromatography mass spectrometry (HS-SPME-GC-MS). The 31 volatile compounds analysed include ethyl esters, acetates, acids and alcohols. Although these compounds have a range of functional groups, chemical properties, volatilities, affinities for the SPME fibre, and are found in wine at various concentrations, the accuracy of the analysis was achieved with the use of polydeuterated internal standards for stable isotope dilution analyses (SIDA). Nine of the labelled standards were commercially available, while 22 were synthesised. The method was validated by a series of duplicate spiked standard additions to model, white and red wine matrices over the concentration range relevant for each compound in wine. This demonstrated that the appropriate use of SIDA helped to account for matrix effects, for instance potential sources of variation such as the relative response to the MS detector, ionic strength, ethanol content and pH of different wine matrices. The resultant calibration functions had correlation coefficients (R²) ranging from 0.995 to 1.000. Each compound could be quantified at levels below its aroma threshold in wine. Relative standard deviations were all <5%. The method was optimised for the best compromise (over the 31 compounds) of wine dilution factor, level of sodium chloride addition, SPME fibre, SPME temperature, SPME time, GC column and MS conditions. Confirmation of identity was achieved by retention time and peak shape, and measurement of at least three ions for each analyte and internal standard with the MS operating in selected ion monitoring mode to facilitate more precise quantitation with a high sampling rate. The method is a valuable research tool with many relevant applications. A novel method for the combined chiral separation and SIDA quantification of 2- and 3-methylbutanoic acid is also demonstrated.     

Analytical performance of three commonly used extraction methods for the gas chromatography–mass spectrometry analysis of wine volatile compounds

The analytical performance of three extraction procedures based on cold liquid–liquid extraction using dicloromethane (LLE), solid phase extraction (SPE) using a styrene–divinylbenzene copolymer and headspace solid phase microextraction (SPME) using a carboxen–polydimethylsiloxane coated fibre has been evaluated based on the analysis of 30 representative wine volatile compounds. From the comparison of the three procedures, LLE and SPE showed very good linearity covering a wide range of concentrations of wine volatile compounds, low detection limits, high recovery for most of the volatile compounds under study and higher sensitivity compared to the headspace-SPME procedure. The latter showed in general, poor recovery for polar volatile compounds. Despite some drawbacks associated with the LLE and SPE procedures such as the more tedious sampling treatment and the use of organic solvents, the analytical performance of both procedures showed that they are more adequate for the analysis of wine volatiles.     

Optimization and validation of headspace sorptive extraction for the analysis of volatile compounds in wine vinegars

Quantification of aroma compounds in wine vinegars is challenging due to the complexity of the matrix and the low concentrations expected. A method for the determination of volatile compounds in wine vinegars employing headspace sorptive extraction-thermal desorption-gas chromatography-mass spectrometry (HSSE-TD-GC-MS) was developed. A central composite design was used to optimize the sampling condition. The proposed method was successfully validated and low detection and quantification limits was obtained. The application of the proposed methodology allows the determination of 53 compounds in different wine vinegars (red, Sherry). Five of them have been detected in wine vinegars for the first time.     

Optimisation of headspace solid-phase microextraction for the analysis of volatile phenols in wine

Headspace solid-phase microextraction has been applied to the analysis of volatile phenols in wine. Silica fibre coated with Carbowax-divinylbenzene was found to be more efficient at extracting these compounds than other fibres such as those coated with polydimethylsiloxane, polyacrylate, carboxen-polydimethylsiloxane, and polydimethylsiloxane-divinylbenzene. Different parameters such as extraction time, temperature of the sample during the extraction, ionic strength and sample volume were optimised using a two-level factorial design expanded further to a central composite design, in order to evaluate several possibly influential and/or interacting factors. The headspace (HS)-SPME procedure developed shows adequate detection and quantitation limits, and linear ranges for correctly analysing these compounds in wine. The recoveries obtained were close to 100%, with repeatability values lower than 16%. The method was applied to a variety of white and red wines.     

Analysis of several phenolic compounds with potential antioxidant properties in grape extracts and wines by high-performance liquid chromatography-photodiode array detection without sample preparation

A RP-HPLC method that allows the separation of several types of phenolic compounds present in grapes and wines by direct injection of samples, using a binary gradient with solvents free of salts and photodiode array detection is described. Results show that more than 15 different phenolic molecules with antioxidant properties (flavan-3-ols, anthocyanins, cinnamic acid derivatives, flavonol derivatives and trans-resveratrol) may be separated in a single run by direct injection of red wine. The method is also valuable for the analysis of these compounds in white wine and in skins, seeds and pulp extracts of red and white grapes.     

Vinegar classification based on feature extraction and selection from headspace solid-phase microextraction/gas chromatography volatile analyses: a feasibility study

Headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography (GC) and multivariate data analysis were applied to classify different vinegar types (white and red, balsamic, sherry and cider vinegars) on the basis of their volatile composition. The collected chromatographic signals were analysed using the stepwise linear discriminant analysis (SLDA) method, thus simultaneously performing feature selection and classification. Several options, more or less restrictive according to the final number of considered categories, were explored in order to identify the one that afforded highest discrimination ability. The simplicity and effectiveness of the classification methodology proposed in the present study (all the samples were correctly classified and predicted by cross-validation) are promising and encourage the feasibility of using a similar strategy to evaluate the quality and origin of vinegar samples in a reliable, fast, reproducible and cost-efficient way in routine applications. The high quality results obtained were even more remarkable considering the reduced number of discriminant variables finally selected by the stepwise procedure. The use of only 14 peaks enabled differentiation between cider, balsamic, sherry and wine vinegars, whereas only 3 variables were selected to discriminate between red (RW) and white wine (WW) vinegars. The subsequent identification by gas chromatography-mass spectrometry (GC-MS) of the volatile compounds associated with the discriminant peaks selected in the classification process served to interpret their chemical significance.     

HPLC determination of volatile phenols in wines

Summary An alternative to the traditional solvent extraction method used to extract and rapidly quantify ethyl-and vinylphenol and ethyl-and vinylgaiacol from wine is presented. The method is based on retention of volatile phenols on adsorbants. Among the tested resins, the most efficient, AG 2-X8 (anion exchange resin), worked as well with a synthetic solution as with wines. The percolation of clarified wine adjusted to pH 9 on this resin permits, in particular, the elimination of organic acids. Phenols are not eluted after rinsing the column with 1N HCl, but are eluted with methanol after this treatment. Good recovery (91 %) and good repeatability are observed. The eluate is directly analysed by HPLC on an RP18 column after two-fold dilution in water. The four volatile phenols were completely separated and detected by UV at 280 nm with high sensitivity (20–40 ppb). No interference with other compounds were noted in the different wines analysed.     

DCMC as a Promising Alternative to Bentonite in White Wine Stabilization. Impact on Protein Stability and Wine Aromatic Fraction

Protein haze in white wine is one of the most common non-microbial defects of commercial wines, with bentonite being the main solution utilized by the winemaking industry to tackle this problem. Bentonite presents some serious disadvantages, and several alternatives have been proposed. Here, an alternative based on a new cellulose derivative (dicarboxymethyl cellulose, DCMC) is proposed. To determine the efficiency of DCMC as a bentonite alternative, three monovarietal wines were characterized, and their protein instability and content determined by a heat stability test (HST) and the Bradford method, respectively. The wines were treated with DCMC to achieve stable wines, as shown by the HST, and the efficacy of the treatments was assessed by determining, before and after treatment, the wine content in protein, phenolic compounds, sodium, calcium, and volatile organic compounds (VOCs) as well as the wine pH. DCMC applied at dosages such as those commonly employed for bentonite was able to reduce the protein content in all tested wines and to stabilize all but the Moscatel de Setúbal varietal wine. In general, DCMC was shown to induce lower changes in the wine pH and phenolic content than bentonite, reducing the wine calcium content. Regarding which VOCs are concerned, DCMC produced a general impact similar to that of bentonite, with differences depending on wine variety. The results obtained suggest that DCMC can be a sustainable alternative to bentonite in protein white wine stabilization.     

Determination of wine aroma compounds by dehydration followed by GC/MS

A new extraction method for the analysis of the volatile fraction of white and red wines has been developed and validated. A dehydration step with MgSO4 separated an aroma compounds-rich alcoholic-glycerine layer. Spiked samples showed good recoveries in the range between 75 and 120% with CVmax% of 17, except for 2-phenylethanol and y-butyrolactone for which recoveries in red wines were under 60%, and for monoethylsuccinate, where recoveries averaged 50 and 60% in white and red wines, respectively. Method repeatability and intermediate precision showed good CVmax% with minimum and maximum values between 7.7 and 24, and between 18.7 and 25.0, respectively. The average determination coefficients were greater than 0.99 with CVmax% of 13. The instrumental LOD and LOQ were, in all cases, under 0.05 mg/L, except for 2,3 butanediol (0.20 mg/L). Overall, the presence of wine matrix affected aroma compounds responses in GC/MS analysis. These observations indicate that the use of a matrix-matched calibration curve is mandatory to obtain reliable quantitative data.     

Assessment of the matrix effect on the headspace solid-phase microextraction (HS-SPME) analysis of chlorophenols in wines

In this paper, we propose a comparative study to check the matrix effect on the extraction of three chlorophenols, 2,4,6-trichlorophenol (TCP), 2,3,4,6-tetrachlorophenol and pentachlorophenol, direct precursors of 2,4,6-trichloroanisole, in synthetic and commercial wines (white and red wines). A rapid, simple and sensitive methodology based on solid-phase microextraction (SPME) and GC with electron capture detection (GC-ECD) and mass spectrometric detection (GC-MS) was developed and the variables affecting the extraction process (temperature, time and salt content) were examined employing a factorial design at two levels. Since GC-ECD does not allow the clear identification of target analytes in white wine, owing to overlapped interferences, GC-MS/MS was used for subsequent examinations. Calibration curves were constructed in synthetic, white and red wine. Significant differences between the slopes of synthetic and red wine, with the exception of TCP, were observed. Analytical parameters were evaluated and satisfactory results were obtained, showing the usefulness of the headspace SPME (HS-SPME) method for determining chlorophenolic compounds in wines.     

Headspace solid-phase microextraction of sulphides and disulphides using Carboxen-polydimethylsiloxane fibers in the analysis of wine aroma

Headspace solid-phase microextraction was applied to gas chromatography coupled to flame photometric detection to develop a method for analysing volatile sulphides and disulphides in wine. The Carboxen-polydimethylsiloxane-coated silica fiber was tested and different parameters such as presampling time, ionic strength, stirring, headspace volume, ethanol concentration, time and temperature of extraction were optimized to make extraction as efficient as possible. The optimized conditions enabled limits of detection to be obtained at the ng/l levels. The fiber tested has a strong affinity for the sulphur compounds studied and enables these analytes to be quantitatively determined in wines. The Carboxen-polydimethylsiloxane-coated fiber is more efficient at extracting than fibers such as those which are polydimethylsiloxane-coated and polyacrylate-coated, but its repeatability is worse. The overall process was successfully applied to identify and quantify sulphur compounds in white, red, rose and vintage wines.     

Improved method to quantitatively determine powerful odorant volatile thiols in wine by headspace solid-phase microextraction after derivatization

A solid-phase microextraction (SMPE) method coupled to a gas chromatography–mass spectrometry analysis was optimized to analyze the pentafluorobenzyl bromide (PFBBr) derivatives of the volatile thiols 4-methyl-4-mercapto-2-pentanone (4MMP), 3-mercaptohexyl acetate (3MHA) and 3-mercaptohexanol (3MH) in wine. This method used deuterated analogue compounds as internal standards. It allowed us to significantly reduce the matrix effect, resulted in good repeatability for all the compounds (RDS < 9.5% for 4MMP; <6% for 3MH and <4.1% for 3MHA) with limits of detection below their odour thresholds. The method was validated using white, rosé and red wines. When applied to analyze different wines, quantities closed to the odour threshold were determined.     

Ochratoxin A survey in Portuguese wine by LC-FD with direct injection

Wine and grape juices were identified as one of the most important sources of ochratoxin A (OTA), a mycotoxin with diverse toxic effects that naturally appears in food and foodstuffs all over the world.The aim of this study was to assess the OTA levels in Portuguese wines through the application of a simple and accurate method based on liquid chromatography (LC) with direct injection, followed by fluorescence detection (FD).Randomly selected wine samples were used to evaluate the performance of direct injection as efficient, fast, inexpensive and safe sample preparation method. The proposed method was successfully validated. The limit of quantification (LOQ) was 1.0 μg/L and OTA recoveries from wine samples, spiked at the three fortification levels, were higher than 85.4%, with RSDs lower than 9.6% for both red and white wines. The presence of OTA was confirmed by methyl ester derivatization followed by LC analysis.Data on OTA levels were obtained for 60 Portuguese red and white wine samples. OTA was found in 12 samples, nine (26%) red wine samples and three (12%) white wine samples. Only one red wine sample and one white wine sample presented a contamination level above the LOQ, with 1.23 and 2.4 μg/L, respectively. It should be pointed out that this white wine sample exceeded the EC maximum permitted level of 2.0 μg/L. The safe dose established as 120 ng/kg body weight/week was not exceeded by the weekly intake estimated for the samples contaminated above the LOQ.     

Beyond the characterization of wine aroma compounds: looking for analytical approaches in trying to understand aroma perception during wine consumption

The volatile compounds present in wines are responsible for the quality of the wine aroma. The analysis of these compounds requires different analytical techniques depending on the type of compounds and their concentration. The importance at sensorial level of each compound should be evaluated by using olfactometric techniques and reconstitution and omission studies. In addition, wine aroma is influenced by other factors such as wine matrix, which could affect the compounds’ volatility, decreasing or increasing their concentration in the headspace above the wine. Moreover, when a wine is consumed, several oral physiological variables could affect aroma perception. The focus of this review is to outline the most recent advances in wine aroma analysis and the most innovative techniques in trying to elucidate the main factors that influence wine aroma perception during consumption.     

Effect of maturation in small oak casks on the volatility of red wine aroma compounds

Wood maturation of red wine produces complex interactions between oak extracted compounds and pre-existing components in wine. Wood contributes with some aroma volatiles; but the whole volatile fraction, including the volatiles extracted from grape or produced during fermentation, could be involved in interactions with the non-volatile fraction. Samples of red wine with increasing time of wood contact, matured in 25 l casks of new American oak, were analysed on their volatile composition both in solution and headspace, and phenolic components. There was an increase of acetic acid and acetate esters as the time of wood contact increased. On the other hand, ethyl esters decreased as the time in wood increased. Some volatiles showed different behaviour when comparing their relationship between headspace and solution from two different casks, even though they were treated as replicates. This different evolution among casks was also consistent with different evolution of phenolic compounds, suggesting that there was an interaction among the volatile fraction and phenolic components.     

Characterisation of the flavour profile from Graciano Vitis vinifera wine variety by a novel dual stir bar sorptive extraction methodology coupled to thermal desorption and gas chromatography–mass spectrometry

The aim of this work was to develop a new analytical technique for the study of the organoleptic compounds (flavour profile) of the Graciano Vitis vinifera wine variety. The cv. Graciano is a singular variety of red grapes with its origins in La Rioja and Navarra (northern Spain). This variety transfers an intense red colour, aroma and high acidity to musts and provides greater longevity and, consequently, a better capacity for ageing wine. A new dual-stir bar sorptive extraction approach coupled with thermal desorption (TD) and GC–MS has been used to extract the volatile and semivolatile compounds. In this extraction step, the optimal values for the experimental variables were obtained through the Response Surface Methodology (RSM). Full scan chromatogram data were evaluated with two deconvolution software tools, and the results were compared. The volatile and semivolatile components were identified with an MS match ≥80%. As a result, the flavour metabolome of the Graciano Vitis vinifera wine variety was obtained, and 205 metabolites were identified using different databases. These metabolites were grouped into esters, acids, alcohols, nitrogen compounds, furans, lactones, ketones, aldehydes, phenols, terpenes, norisoprenoids, sulphur compounds, acetals and pyrans. The majority of the metabolites observed had already been reported in the literature; however, this work also identified new, previously unreported metabolites in red wines, which may be characteristic of the Graciano variety.     

Determination of ochratoxin A in wine and beer by immunoaffinity column cleanup and liquid chromatographic analysis with fluorometric detection: collaborative study.

The accuracy, repeatability, and reproducibility characteristics of a liquid chromatographic method for the determination of ochratoxin A (OTA) in white wine, red wine, and beer were established in a collaborative study involving 18 laboratories in 10 countries. Blind duplicates of blank, spiked, and naturally contaminated materials at levels ranging from < or =0.01 to 3.00 ng/mL were analyzed. Wine and beer samples were diluted with a solution containing polyethylene glycol and sodium hydrogen carbonate, and the diluted samples were filtered and cleaned up on an immunoaffinity column. OTA was eluted with methanol and quantified by reversed-phase liquid chromatography with fluorometric detection. Average recoveries from white wine, red wine, and beer ranged from 88.2 to 105.4% (at spiking levels ranging from 0.1 to 2.0 ng/mL), from 84.3 to 93.1% (at spiking levels ranging from 0.2 to 3.0 ng/mL), and from 87.0 to 95.0% (at spiking levels ranging from 0.2 to 1.5 ng/mL), respectively. Relative standard deviations for within-laboratory repeatability (RSDr) ranged from 6.6 to 10.8% for white wine, from 6.5 to 10.8% for red wine, and from 4.7 to 16.5% for beer. Relative standard deviations for between-laboratories reproducibility (RSDR) ranged from 13.1 to 15.9% for white wine, from 11.9 to 13.6% for red wine, and from 15.2 to 26.1% for beer. HORRAT values were < or =0.4 for the 3 matrixes.     

Microextraction of volatile compounds from wine samples and their determination by GC-FID. The effect of the salts and extraction solvents used

Summary This study describes the optimization of microextraction as a method for extracting volatile compounds from wine. The study has been applied to twelve compounds present in wine: 3-methylbutyl acetate, 3-methyl-1-butanol, ethyl hexanoate, hexanol, ethyl octanoate, butanoic acid, ethyl decanoate, diethyl succinate, hexanoic acid, phenylethanol, octanoic acid, and decanoic acid. These compounds were selected from those identified by GC-MS analysis of a real wine sample. By means of a synthetic wine, the study investigated the influence on extraction yield of the solvents and salts used, the proportion of salts, and the agitation time. The determination was performed by GC with flame ionization detection and an internal standard was used for quantification. The method was applied to samples of white wine from La Rioja.